Elderly Rheumatoid Arthritis Cold/heat Syndrome Of TCM And Knee Cartilage LncRNA Expression Profiling

Objective1.Through the system literature review,summarized the epidemiology of senile rheumatoid arthritis,pathogenesis of Western medicine,clinical features,etiology and pathogenesis of traditional Chinese medicine,TCM syndrome differentiation,the latest research progress.Objective to investigate the application of LncRNA microarray technology in the study of elderly patients with rheumatoid arthritis.2.Through the investigation of elderly patients with rheumatoid arthritis in epidemiology,clinical characteristics,TCM syndrome characteristics of the situation,the correlation between the disease activity and severity of disease,extra articular manifestations,complications,laboratory indexes and TCM type of cold and heat syndrome,provide an objective basis for elderly rheumatoid arthritis syndrome,syndrome differentiation and provide accurate effective method.3.Through the detection of expression of elderly patients with rheumatoid arthritis of knee joint cartilage of long chain non encoding RNA,to explore the differential gene expression in patients with rheumatoid arthritis and osteoarthritis of the knee,looking for possible pathogenesis related target gene;to explore the relationship between genetic differences and TCM cold and heat arthromyodynia,looking for the objective syndrome differentiation standard.For the Chinese medicine to provide an effective method for accurate differentiation.Methods1.Literature research:search elderly rheumatoid arthritis related literature,an overview of the pathogenesis of senile rheumatoid arthritis,to explore the possibility of articular cartilage lesions as immune antigen onset rheumatoid arthritis and LncRNA in patients with rheumatoid arthritis research;research on TCM etiology and syndrome differentiation and epidemiological review of elderly rheumatoid arthritis,analysis the status quo of elderly rheumatoid arthritis syndromes objectification.2.Clinical research:review and analysis of.the Department of traditional Chinese medicine of Genenral Hospital of PLA Guangzhou Military Area/clinic and ward of Rheumatology,150 cases of rheumatoid arthritis patients,with onset age over 60 years old were divided into RA group and youth group RA,compared two groups of patients with gender ratio,disease activity,disease severity and extra articular manifestations,complications,laboratory index clinical observation;Study on 60 cases of elderly patients with RA:cold aggravated,Wen Zeshu is cold and dampness;joint swelling heat pain,cold and cool for pain reduction in rheumatic fever.Clinical observation,including the basic situation of the TCM syndrome characteristics,disease activity,disease severity and extra articular manifestations,complications,laboratory indicators,the use of modern statistical methods,to explore the relationship between these indexes and cold/heat arthromyodynia.3.Experimental study:to get 15 cases of knee cartilage.The patients were divided into 7 groups:RA group,KOA group and normal group,with a total of 3 cases in the elderly group.Total RNA was extracted from cartilage tissue,using LncRNA+mRNA microarray,comparison of elderly RA group and KOA group,lncRNA mRNA expression,while detecting RA in elderly group cold/heat syndrome between IncRNA and mRNA expressiona preliminary analysis of differential gene function using GO and KEGG analysis method.Using the method of gene co expression and the prediction of target genes,we can get the key IncRNAs of aged RA.Screening and cold/heat arthralgia arthritis immune related genes,established by computer analysis of elderly patients with rheumatoid arthritis of cold/heat arthralgia syndrome related gene expression profiles reveal cold/heat arthromyodynia when forming all related genes and their functions,the essence of expression from the whole gene level to elucidate the syndromes,and provide scientific basis for the elderly RA precision of TCM syndrome differentiation and treatment.Results1.Clinical research:The results showed that the score of DAS28,ACR20 and ACR50 and the effective rate of DAS28 in both treatment groups were significantly higher than that in control group after treated for 4 weeks（P<0.05）.After treated for 12 weeks,the total effective rate of Han Bi group,Re Bi group and the control group total was 88%,80.0%and 77.6%,respectively;and the efficiency rate was 56%,53.1%and 38.8%,respectively.There was a significant difference between week 12 and week 4 for all of the three groups（P<0.05）.The VAS score and the doctor/patient VAS score in both treatment groups were significantly higher than that in the MTX group after treated for 4 weeks with a better general condition（P<0.05）.Han Bi group had a higher VAS score and the doctor/patient VAS score,more swollen joints,stronger grips of both left and right hand and more painful joints than Re Bi group and the MTX group（P<0.05）.There was no significant difference in the duration of morning stiffness and HAQ improvement for all of the three groups.Han Bi group had a better general condition like temperature,thirst,sleep,stool and urine condition compared with before.The therapeutic efficacy of Han Bi group were significantly better than Re Bi group and the control group（P<0.05）.The improvement of general condition for Re Bi group was not obvious.There was no significant difference in the incidence of adverse events for the three groups.2.Experimental study:Gene expression differences were obtained for the samples of both groups.The number of significantly differential expressed genes for the samples of both groups were over 3000,1683 up-regulated genes,2540 down-regulated genes and 95 unknown genes,respectively.Molecular and biological function analysis showed that down-regulated genes were primarily involved in cell apoptosis,proliferation,immune response,cell metabolism,cell communication,bone metabolism,hormones,cell signal transduction, inflammation,protein synthesis translation.3.The experimental study found that the elderly RA group compared with the KOA group,screened the genes of the lncRNA difference expression to be 5862（27.35%）,of which 3338 genes were raised,and 2524 genes were lowered.The differences in mRNAs were 3343（26.68%）,including 1787 genes and 1556 genes.These genes are likely to be the target genes for geriatric RA.The older RA group and the heat group screened 317（1.18%）of the differences in the expression of the IncRNAs,with the increase of 263 genes and 54 genes.148（0.94 percent）of the differences were expressed in mRNAs,with the increase in gene 126,and the reduction of 22 genes.These genes are likely to be different targets for geriatric RA.Through KEGG RA for the elderly and KOA group differences in expression of mRNA analysis,related pathways have phosphatidyl inositol signal system,glucagon signaling pathways,cGMP-PKG signaling pathways,N-h biosynthesis pathway,phosphatidyl inositol signal system,etc.4.Elderly RA "cold/hot bi syndrome" related gene prediction:elderly RA was winter and heat bi group to screen differentially expressed（1.18%）of 317 differentially expressed IncRNAs,which raised 263 genes,cut article 54 genes;148（0.94 percent）of the differences were expressed in mRNAs,with the increase in gene 126,and the reduction of 22 genes.These genes are likely to be the target genes for the geriatric syndromes of the geriatric RA.The differences genes were screened for the elderly RA group and the thermal group:G056613,LOC285629,G005949,G008220;The gene for the difference between the elderly RA and the thermal bi group was screened:XLOC₀02287,G082016,AC092667.2,G065878.The differences between the elderly RA groups and the thermal groups were screened out.The differences in the reduction of the elderly RA group and the thermal bi group were selected.There is a small variation in the expression of the lncRNA and mRNA in geriatric RA.Conclusion1.LncRNA in elderly rheumatoid arthritis research progress of analysis of elderly rheumatoid arthritis research status quo of TCM syndrome types objective.2.Elderly RA heat bi syndrome and blood sedimentation,c-reactive protein and TCM symptoms correlated with integral（P<0.05）,the group than was winter certification of hot blood sedimentation,high c-reactive protein and TCM symptom integral;Elderly RA,deadlocked of morning of cold/heat bi syndrome of traditional Chinese medicine and symptoms may have a certain correlation（P<0.1）,while no significant under 5%significance level,but close to 10%,hot bi group than cold bi group are more likely to have the symptoms of morning stiffness;Cold/hot bi syndrome with BMI,age,course of the disease in elderly patients with RA,rheumatoid factor,DAS28,CDAI,VAS score,the number of swollen joints,the tenderness of joint number correlation is not obvious.Study found that older RA TCM cold/hot bi syndrome and gender,the initial joint three,rheumatoid disease,diabetes,high blood pressure,high uric acid has no obvious relation,hyperlipidemia,coronary heart disease（CHD）.3.In the research of cartilage LncRNA elderly RA group compared with KOA group,chip of gene expression that elderly RA group is similar to the characteristics of the tumor tissue,such as oxidative phosphorylation,arachidonic acid metabolism,acyl inositol phosphate signaling system,etc.They can make the inflammatory cells,synovial hyperplasia,immune network unbalance leads to degradation and destruction of bone and cartilage,synovial cell and fibroblasts class sample tumor growth and secretion of promoting inflammatory cytokines and chemokines and metalloproteinases.In addition,elderly RA group and KOA way comparison,chip tip energy metabolism and RA disease have higher correlation:such as other polysaccharide degradation,N-h biosynthesis,glucagon signaling pathways,polysaccharide,biosynthesis,synthesis and secretion of aldosterone signaling pathways,etc.They can affect the activity of glycolysis in the tissue of RA,resulting in hypoxia and acidic microenvironment.Glycolytic pathway of key enzymes such as hexose phosphate isomerase,enolization enzyme,aldolase and triose phosphate isomerase as an antigen,the induced by RA also play a role in immune response;Lactic acid and pyruvate affect the metabolism of the RA synovial matrix,stimulating the proliferation of abnormal cells,the formation of blood vessels and vasculature.4.The elderly RA "cold/hot bi syndrome" the related gene prediction:G056613,LOC285629,G005949,G008220,XLOC₀02287,G082016,AC092667,G065878,TCRBV6S6A2T,SHB,CEP104,OR2W3,ARHGAP39,BST1.Elderly RA cold/hot bi syndrome syndrome between the comparison,there are a few differences IncRNA,mRNA expression.Gene expression level to clarify the essence of syndrome,RA for your old age will provide a scientific basis for precision of traditional Chinese medicine syndrome differentiation and treatment.